Affiliation:
1. Centro de Biotecnología y Genómica de Plantas (CBGP) (UPM-INIA/CSIC), Autopista M40, km 38, Campus Montegancedo, Pozuelo de Alarcón, 28223 Madrid, Spain
Abstract
Plant viral nanoparticles (VNPs) have become an attractive platform for the development of novel nanotools in the last years because of their safety, inexpensive production, and straightforward functionalization. Turnip mosaic virus (TuMV) is one example of a plant-based VNP used as a nanobiotechnological platform either as virions or as virus-like particles (VLPs). Their functionalization mainly consists of coating their surface with the molecules of interest via chemical conjugation or genetic fusion. However, because of their limitations, these two methods sometimes result in non-viable constructs. In this paper, we applied the SpyTag/SpyCatcher technology as an alternative for the functionalization of TuMV VLPs with peptides and proteins. We chose as molecules of interest the green fluorescent protein (GFP) because of its good traceability, as well as the vasoactive intestinal peptide (VIP), given the previous unsuccessful attempts to functionalize TuMV VNPs by other methods. The successful conjugation of VLPs to GFP and VIP using SpyTag/SpyCatcher was confirmed through Western blot and electron microscopy. Moreover, the isopeptide bond between SpyTag and SpyCatcher occurred in vivo in co-agroinfiltrated Nicotiana benthamiana plants. These results demonstrated that SpyTag/SpyCatcher improves TuMV functionalization compared with previous approaches, thus implying the expansion of the application of the technology to elongated flexuous VNPs.
Funder
Comunidad de Madrid
INIA
ERANet project
Spanish Ministry of Universities and UCM
Subject
Virology,Infectious Diseases
Reference38 articles.
1. Virus-Based Nanoparticles as Versatile Nanomachines;Koudelka;Annu. Rev. Virol.,2015
2. Synthetic Plant Virology for Nanobiotechnology and Nanomedicine;Steele;Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol.,2017
3. Venkataraman, S., and Hefferon, K. (2021). Application of Plant Viruses in Biotechnology, Medicine, and Human Health. Viruses, 13.
4. Surface Functionalization of Rod-Shaped Viral Particles for Biomedical Applications;Vaidya;ACS Appl. Bio Mater.,2022
5. Plant Viral Elongated Nanoparticles Modified for Log-Increases of Foreign Peptide Immunogenicity and Specific Antibody Detection;Lunello;J. Biotechnol.,2013
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